Filter
Associated Lab
- Ahrens Lab (2) Apply Ahrens Lab filter
- Aso Lab (3) Apply Aso Lab filter
- Baker Lab (1) Apply Baker Lab filter
- Betzig Lab (8) Apply Betzig Lab filter
- Bock Lab (1) Apply Bock Lab filter
- Branson Lab (7) Apply Branson Lab filter
- Card Lab (4) Apply Card Lab filter
- Cardona Lab (8) Apply Cardona Lab filter
- Cui Lab (1) Apply Cui Lab filter
- Dickson Lab (1) Apply Dickson Lab filter
- Druckmann Lab (3) Apply Druckmann Lab filter
- Dudman Lab (4) Apply Dudman Lab filter
- Eddy/Rivas Lab (1) Apply Eddy/Rivas Lab filter
- Feliciano Lab (1) Apply Feliciano Lab filter
- Fetter Lab (4) Apply Fetter Lab filter
- Funke Lab (1) Apply Funke Lab filter
- Gonen Lab (11) Apply Gonen Lab filter
- Grigorieff Lab (6) Apply Grigorieff Lab filter
- Harris Lab (5) Apply Harris Lab filter
- Heberlein Lab (1) Apply Heberlein Lab filter
- Hermundstad Lab (1) Apply Hermundstad Lab filter
- Hess Lab (4) Apply Hess Lab filter
- Jayaraman Lab (4) Apply Jayaraman Lab filter
- Ji Lab (5) Apply Ji Lab filter
- Keleman Lab (1) Apply Keleman Lab filter
- Keller Lab (2) Apply Keller Lab filter
- Lavis Lab (16) Apply Lavis Lab filter
- Lee (Albert) Lab (6) Apply Lee (Albert) Lab filter
- Leonardo Lab (2) Apply Leonardo Lab filter
- Lippincott-Schwartz Lab (9) Apply Lippincott-Schwartz Lab filter
- Liu (Zhe) Lab (5) Apply Liu (Zhe) Lab filter
- Looger Lab (6) Apply Looger Lab filter
- Magee Lab (2) Apply Magee Lab filter
- Menon Lab (1) Apply Menon Lab filter
- Pachitariu Lab (1) Apply Pachitariu Lab filter
- Reiser Lab (6) Apply Reiser Lab filter
- Riddiford Lab (1) Apply Riddiford Lab filter
- Romani Lab (6) Apply Romani Lab filter
- Rubin Lab (15) Apply Rubin Lab filter
- Saalfeld Lab (4) Apply Saalfeld Lab filter
- Scheffer Lab (4) Apply Scheffer Lab filter
- Schreiter Lab (4) Apply Schreiter Lab filter
- Shroff Lab (1) Apply Shroff Lab filter
- Simpson Lab (2) Apply Simpson Lab filter
- Singer Lab (6) Apply Singer Lab filter
- Spruston Lab (1) Apply Spruston Lab filter
- Stern Lab (8) Apply Stern Lab filter
- Sternson Lab (2) Apply Sternson Lab filter
- Svoboda Lab (9) Apply Svoboda Lab filter
- Truman Lab (6) Apply Truman Lab filter
- Turaga Lab (3) Apply Turaga Lab filter
- Turner Lab (2) Apply Turner Lab filter
- Wu Lab (1) Apply Wu Lab filter
- Zlatic Lab (7) Apply Zlatic Lab filter
Associated Project Team
- Fly Descending Interneuron (1) Apply Fly Descending Interneuron filter
- Fly Functional Connectome (4) Apply Fly Functional Connectome filter
- Fly Olympiad (1) Apply Fly Olympiad filter
- FlyEM (4) Apply FlyEM filter
- FlyLight (2) Apply FlyLight filter
- GENIE (3) Apply GENIE filter
- ThalamoSeq (1) Apply ThalamoSeq filter
- Tool Translation Team (T3) (3) Apply Tool Translation Team (T3) filter
- Transcription Imaging (6) Apply Transcription Imaging filter
Associated Support Team
- Anatomy and Histology (2) Apply Anatomy and Histology filter
- Cryo-Electron Microscopy (4) Apply Cryo-Electron Microscopy filter
- Electron Microscopy (1) Apply Electron Microscopy filter
- Fly Facility (1) Apply Fly Facility filter
- Integrative Imaging (1) Apply Integrative Imaging filter
- Project Technical Resources (1) Apply Project Technical Resources filter
- Quantitative Genomics (2) Apply Quantitative Genomics filter
- Scientific Computing Software (9) Apply Scientific Computing Software filter
- Viral Tools (1) Apply Viral Tools filter
- Vivarium (1) Apply Vivarium filter
Publication Date
- December 2017 (15) Apply December 2017 filter
- November 2017 (11) Apply November 2017 filter
- October 2017 (7) Apply October 2017 filter
- September 2017 (14) Apply September 2017 filter
- August 2017 (15) Apply August 2017 filter
- July 2017 (20) Apply July 2017 filter
- June 2017 (18) Apply June 2017 filter
- May 2017 (25) Apply May 2017 filter
- April 2017 (21) Apply April 2017 filter
- March 2017 (15) Apply March 2017 filter
- February 2017 (7) Apply February 2017 filter
- January 2017 (18) Apply January 2017 filter
- Remove 2017 filter 2017
186 Janelia Publications
Showing 31-40 of 186 resultsCalcium carbonate platforms produced by reef-building stony corals over geologic time are pervasive features around the world [1]; however, the mechanism by which these organisms produce the mineral is poorly understood (see review by [2]). It is generally assumed that stony corals precipitate calcium carbonate extracellularly as aragonite in a calcifying medium between the calicoblastic ectoderm and pre-existing skeleton, separated from the overlying seawater [2]. The calicoblastic ectoderm produces extracellular matrix (ECM) proteins, secreted to the calcifying medium [3-6], which appear to provide the nucleation, alteration, elongation, and inhibition mechanisms of the biomineral [7] and remain occluded and preserved in the skeleton [8-10]. Here we show in cell cultures of the stony coral Stylophora pistillata that calcium is concentrated in intracellular pockets that are subsequently exported from the cell where a nucleation process leads to the formation of extracellular aragonite crystals. Analysis of the growing crystals by lattice light-sheet microscopy suggests that the crystals elongate from the cells' surfaces outward.
The ability to reproducibly target expression of transgenes to small, defined subsets of cells is a key experimental tool for understanding many biological processes. The Drosophila nervous system contains thousands of distinct cell types and it has generally not been possible to limit expression to one or a few cell types when using a single segment of genomic DNA as an enhancer to drive expression. Intersectional methods, in which expression of the transgene only occurs where two different enhancers overlap in their expression patterns, can be used to achieve the desired specificity. This report describes a set of over 2,800 transgenic lines for use with the split-GAL4 intersectional method.
Histone CENP-A-containing nucleosomes play an important role in nucleating kinetochores at centromeres for chromosome segregation. However, the molecular mechanisms by which CENP-A nucleosomes engage with kinetochore proteins are not well understood. Here, we report the finding of a new function for the budding yeast Cse4/CENP-A histone-fold domain interacting with inner kinetochore protein Mif2/CENP-C. Strikingly, we also discovered that AT-rich centromere DNA has an important role for Mif2 recruitment. Mif2 contacts one side of the nucleosome dyad, engaging with both Cse4 residues and AT-rich nucleosomal DNA. Both interactions are directed by a contiguous DNA- and histone-binding domain (DHBD) harboring the conserved CENP-C motif, an AT hook, and RK clusters (clusters enriched for arginine-lysine residues). Human CENP-C has two related DHBDs that bind preferentially to DNA sequences of higher AT content. Our findings suggest that a DNA composition-based mechanism together with residues characteristic for the CENP-A histone variant contribute to the specification of centromere identity.
Leukocytes and other amoeboid cells change shape as they move, forming highly dynamic, actin-filled pseudopods. Although we understand much about the architecture and dynamics of thin lamellipodia made by slow-moving cells on flat surfaces, conventional light microscopy lacks the spatial and temporal resolution required to track complex pseudopods of cells moving in three dimensions. We therefore employed lattice light sheet microscopy to perform three-dimensional, time-lapse imaging of neutrophil-like HL-60 cells crawling through collagen matrices. To analyze three-dimensional pseudopods we: (i) developed fluorescent probe combinations that distinguish cortical actin from dynamic, pseudopod-forming actin networks, and (ii) adapted molecular visualization tools from structural biology to render and analyze complex cell surfaces. Surprisingly, three-dimensional pseudopods turn out to be composed of thin (<0.75 µm), flat sheets that sometimes interleave to form rosettes. Their laminar nature is not templated by an external surface, but likely reflects a linear arrangement of regulatory molecules. Although we find that Arp2/3-dependent pseudopods are dispensable for three-dimensional locomotion, their elimination dramatically decreases the frequency of cell turning, and pseudopod dynamics increase when cells change direction, highlighting the important role pseudopods play in pathfinding.
Cells alter their mechanical properties in response to their local microenvironment; this plays a role in determining cell function and can even influence stem cell fate. Here, we identify a robust and unified relationship between cell stiffness and cell volume. As a cell spreads on a substrate, its volume decreases, while its stiffness concomitantly increases. We find that both cortical and cytoplasmic cell stiffness scale with volume for numerous perturbations, including varying substrate stiffness, cell spread area, and external osmotic pressure. The reduction of cell volume is a result of water efflux, which leads to a corresponding increase in intracellular molecular crowding. Furthermore, we find that changes in cell volume, and hence stiffness, alter stem-cell differentiation, regardless of the method by which these are induced. These observations reveal a surprising, previously unidentified relationship between cell stiffness and cell volume that strongly influences cell biology.
Labeled probes, and methods of use thereof, comprise a Cas polypeptide conjugated to gRNA that is specific for target nucleic acid sequences, including genomic DNA sequences. The probes and methods can be used to label nucleic acid sequences without global DNA denaturation. The presently-disclosed subject matter meets some or all of the above identified needs, as will become evident to those of ordinary skill in the art after a study of information provided in this document.
View Publication PageTo ensure disjunction to opposite poles during anaphase, sister chromatids must be held together following DNA replication. This is mediated by cohesin, which is thought to entrap sister DNAs inside a tripartite ring composed of its Smc and kleisin (Scc1) subunits. How such structures are created during S phase is poorly understood, in particular whether they are derived from complexes that had entrapped DNAs prior to replication. To address this, we used selective photobleaching to determine whether cohesin associated with chromatin in G1 persists in situ after replication. We developed a non-fluorescent HaloTag ligand to discriminate the fluorescence recovery signal from labeling of newly synthesized Halo-tagged Scc1 protein (pulse-chase or pcFRAP). In cells where cohesin turnover is inactivated by deletion of WAPL, Scc1 can remain associated with chromatin throughout S phase. These findings suggest that cohesion might be generated by cohesin that is already bound to un-replicated DNA.
The development of genetically encoded self-labeling protein tags such as the HaloTag and SNAP-tag has expanded the utility of chemical dyes in microscopy. Intracellular labeling using these systems requires small, cell-permeable dyes with high brightness and photostability. We recently discovered a general method to improve the properties of classic fluorophores by replacing N,N-dimethylamino groups with four-membered azetidine rings to create the "Janelia Fluor" dyes. Here, we describe the synthesis of the HaloTag and SNAP-tag ligands of Janelia Fluor 549 and Janelia Fluor 646 as well as standard labeling protocols for use in ensemble and single-molecule cellular imaging.
Developmental genes can have complex cis-regulatory regions, with multiple enhancers scattered across stretches of DNA spanning tens or hundreds of kilobases. Early work revealed remarkable modularity of enhancers, where distinct regions of DNA, bound by combinations of transcription factors, drive gene expression in defined spatio-temporal domains. Nevertheless, a few reports have shown that enhancer function may be required in multiple developmental stages, implying that regulatory elements can be pleiotropic. In these cases, it is not clear whether the pleiotropic enhancers employ the same transcription factor binding sites to drive expression at multiple developmental stages or whether enhancers function as chromatin scaffolds, where independent sets of transcription factor binding sites act at different stages. In this work we have studied the activity of the enhancers of the shavenbaby gene throughout D. melanogaster development. We found that all seven shavenbaby enhancers drive gene expression in multiple tissues and developmental stages at varying levels of redundancy. We have explored how this pleiotropy is encoded in two of these enhancers. In one enhancer, the same transcription factor binding sites contribute to embryonic and pupal expression, whereas for a second enhancer, these roles are largely encoded by distinct transcription factor binding sites. Our data suggest that enhancer pleiotropy might be a common feature of cis-regulatory regions of developmental genes and that this pleiotropy can be encoded through multiple genetic architectures.
Neurons relevant to a particular behavior are often widely dispersed across the brain. To record activity in groups of individual neurons that might be distributed across large distances, neuroscientists and optical engineers have been developing a new type of microscope called a mesoscope. Mesoscopes have high spatial resolution and a large field of view. This Q&A will discuss this exciting new technology, highlighting a particular instrument, the two-photon random access mesoscope (2pRAM).